A Transgenic Zebrafish for Monitoring In Vivo Microtubule Structures
A Transgenic Zebrafish for Monitoring In Vivo Microtubule Structures
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DOI：
10.1002/dvdy.22400
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发表时间：
2010-10-01
影响因子：
2.5
通讯作者：
Kawakami, Koichi
中科院分区：
文献类型：
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作者：
Asakawa, Kazuhide;Kawakami, Koichi
微管（MT）细胞骨架通过调节神经元祖细胞的增殖、神经元迁移和轴突导向在脑发育中起着至关重要的作用。然而，用于监测完整脑中MT的方法在脊椎动物中受到限制。在这里，我们报告了一个转基因斑马鱼线监测MT在体内。该报告细胞系携带编码绿色荧光蛋白（GFP）标记的微管蛋白基因的转基因，该基因连接到上游激活序列（UAS），即酵母Gal 4转录激活因子的识别序列。通过将该报告细胞系与适当的转基因Gal 4驱动细胞系杂交，我们在从胚胎阶段到成体阶段的各种细胞类型中诱导GFP标记的微管蛋白。在表达修饰的微管蛋白的幼虫中，单个MT丝和其他MT结构，包括增殖神经元祖细胞中的有丝分裂纺锤体，被清楚地可视化。因此，转基因UAS报告细胞系应可用于直接监测完整脑中的MT。Developmental Dynamics 239：2695-2699，2010. (C)2010 Wiley-Liss，Inc.
The microtubule (MT) cytoskeleton plays crucial roles in brain development by regulating the proliferation of neuronal progenitor cells, neuronal migration and axon guidance. Methods for monitoring MT in the intact brain, however, have been limited in vertebrates. Here, we report a transgenic zebrafish line for monitoring MT in vivo. This reporter line carries a transgene encoding the green fluorescent protein (GFP) -tagged tubulin gene linked to the upstream activating sequence (UAS), the recognition sequence of the yeast Gal4 transcriptional activator. By crossing this reporter line with appropriate transgenic Gal4 driver lines, we induced the GFP-tagged tubulin in various cell types from the embryonic stages to the adult stage. In larvae expressing the modified tubulin, individual MT filaments and other MT structures, including the mitotic spindles in proliferating neuronal progenitor cells, were clearly visualized. Therefore, the transgenic UAS reporter line should be useful for directly monitoring MTs in the intact brain. Developmental Dynamics 239:2695-2699, 2010. (C) 2010 Wiley-Liss, Inc.